Designed Asymmetric Protein Assembly on a Symmetric Scaffold

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2020 Apr 26

PMID 32333708

Citations 4

Authors

Lenne J M Lemmens

Job A L Roodhuizen

Tom F A de Greef

Albert J Markvoort

Luc Brunsveld

Affiliations

Soon will be listed here.

Abstract

Cellular signaling is regulated by the assembly of proteins into higher-order complexes. Bottom-up creation of synthetic protein assemblies, especially asymmetric complexes, is highly challenging. Presented here is the design and implementation of asymmetric assembly of a ternary protein complex facilitated by Rosetta modeling and thermodynamic analysis. The wild-type symmetric CT32-CT32 interface of the 14-3-3-CT32 complex was targeted, ultimately favoring asymmetric assembly on the 14-3-3 scaffold. Biochemical studies, supported by mass-balance models, allowed characterization of the parameters driving asymmetric assembly. Importantly, our work reveals that both the individual binding affinities and cooperativity between the assembling components are crucial when designing higher-order protein complexes. Enzyme complementation on the 14-3-3 scaffold highlighted that interface engineering of a symmetric ternary complex generates asymmetric protein complexes with new functions.

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Designed Asymmetric Protein Assembly on a Symmetric Scaffold.

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PMID: 32333708 PMC: 7383506. DOI: 10.1002/anie.202003626.

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